Abstract:

Monodisperse polystyrene seed particles were prepared by dispersion polymerization of styrene in ethanol and water under a nitrogen atmosphere using benzoyl peroxide as an initiator. Then the monodisperse, porous poly (chloromethylstyrene-co-divinylbenzene) beads in the range of 6-15 μm were prepared by one-step swelling and polymerization method. Based on this media, a new hydrophilic weak cation exchange stationary phase (WCX) for high performance liquid chromatography was synthesized by a new chemical modification method. The stationary phase was evaluated on the behavior of ion exchange, separability, and hydrophilicity. Effects of the nature of salts and pH value on the separation and retention of proteins of the stationary phase were investigated in details. It was found that the WCX resin follows ion exchange chromatographic retention mechanism. It was also used for the rapid separation and purification of lysozyme from egg white in only one step. The purity was found more than 96% and specific bioactivity of the purified lysozyme was as high as 71184 U/mg.

Key words: protein separation , weak cation exchange chromatography, monodisperse porous poly (chloromethylstyrene-co-divinylbenzene) beads